Publications

Summary How cadmium (Cd) tolerance in rice is regulated remains poorly understood. We used a forward genetic approach to investigate Cd tolerance in rice. Using a root elongation assay, we isolated a rice mutant with enhanced Cd tolerance, cadt1 , from an ethyl methanesulphonate (EMS)‐mutagenized population of a widely grown Indica cultivar. The mutant accumulated more Cd in roots but not in shoots and grains. Using genomic resequencing and complementation, we identified OsCADT1 as the causal gene for the mutant phenotype, which encodes a putative serine hydroxymethyltransferase. OsCADT1 protein was localized to the nucleus and the OsCADT1 gene was expressed in both roots and shoots. OsCADT1 mutation resulted in higher sulphur and selenium accumulation in the shoots and grains. Selenate influx in cadt1 was 2.4 times that of the wild‐type. The mutant showed higher expression of the sulphate/selenate transporter gene OsSULTR1;1 and the sulphur‐deficiency‐inducible gene OsSDI1 . Thiol compounds including cysteine, glutathione and phytochelatins were significantly increased in the mutant, underlying its increased Cd tolerance. Growth and grain biomass were little affected. The results suggest that OsCADT1 acts as a negative regulator of sulphate/selenate uptake and assimilation. OsCADT1 mutation increases Cd tolerance and enriches selenium in rice grains, providing a novel solution for selenium biofortification.

Microbial arsenic (As) methylation in paddy soil produces mainly dimethylarsenate (DMA), which can cause physiological straighthead disease in rice. The disease is often highly patchy in the field, but the reasons remain unknown. We investigated within-field spatial variations in straighthead disease severity, As species in rice husks and in soil porewater, microbial composition and abundance of arsM gene encoding arsenite S-adenosylmethionine methyltransferase in two paddy fields. The spatial pattern of disease severity matched those of soil redox potential, arsM gene abundance, porewater DMA concentration, and husk DMA concentration in both fields. Structural equation modelling identified soil redox potential as the key factor affecting arsM gene abundance, consequently impacting porewater DMA and husk DMA concentrations. Core amplicon variants that correlated positively with husk DMA concentration belonged mainly to the phyla of Chloroflexi, Bacillota, Acidobacteriota, Actinobacteriota, and Myxococcota. Meta-omics analyses of soil samples from the disease and non-disease patches identified 5129 arsM gene sequences, with 71% being transcribed. The arsM-carrying hosts were diverse and dominated by anaerobic bacteria. Between 96 and 115 arsM sequences were significantly more expressed in the soil samples from the disease than from the non-disease patch, which were distributed across 18 phyla, especially Acidobacteriota, Bacteroidota, Verrucomicrobiota, Chloroflexota, Pseudomonadota, and Actinomycetota. This study demonstrates that even a small variation in soil redox potential within the anoxic range can cause a large variation in the abundance of As-methylating microorganisms, thus resulting in within-field variation in rice straighthead disease. Raising soil redox potential could be an effective way to prevent straighthead disease.

The interaction of Cd and other heavy metals with soil colloidal particles controls the sequestration, mobility and bioavailability of Cd in soils. In this study, the binding (△G bi ) and adsorption (△G ad ) energies of Cd on colloidal particles of 18 soils were determined by the Wien effect method. The binding energy of Cd on soil colloidal particles varied from 5.3 to 9.9 kJ mol −1 , depending on the soil characteristics including pH, Mn‐oxide content and dissolved organic carbon in the soil. The Cd adsorption energy correlated positively with Mn‐oxide content and pH. In parallel, the extended X‐ray absorption fine structure (EXAFS) spectroscopy was used to determine the speciation of Cd in Cd‐saturated soil samples, which revealed that the outer‐sphere Cd was the predominant species, accounting for 32.2–73.7% of the total adsorbed Cd, and positively correlated to the binding and adsorption energies. Humic acid‐Cd (10.4–42.2%) and montmorillonite‐Cd (2.5–51.2%) were also major species that were identified by EXAFS spectroscopy. The toxicity (log EC 50 ) of Cd in soils to three organisms (earthworm, Collembola and Chinese cabbage) was found to correlate positively with the binding energies, indicating the predictive capability of using binding energies of Cd in different soils as an indicator for evaluating Cd bioavailability and toxicity in soils.

The increasingly widespread usage of silver (Ag) nanoparticles has raised concerns regarding their environmental risk. The behavior of Ag and its transfer risk to the food chain were investigated using a long-term field experiment that commenced in 1942 in which Ag-containing sewage sludge was repeatedly applied to the soil (25 applications during 20 years). The speciation of the Ag in both the sludge and the soils retrieved from the long-term experimental archive was examined using synchrotron-based X-ray absorption spectroscopy, and extractable Ag concentrations from soils were determined using 0.01 M Ca(NO3)2 and 0.005 M DTPA. The total Ag in the sludge during the time period of 1942–1961 ranged from 155 to 463 mg kg–1. These values are 1–2 orders of magnitude higher than those in currently produced sludge (ca. 0.5–20 mg kg–1). Long-term repeated applications of these sludges resulted in an increase of Ag in soils from 1.9 mg kg–1 in the control to up to 51 mg kg–1. The majority (>80%) of the Ag in both the sludge and the sludge-treated soils was present as insoluble Ag2S, thereby markedly reducing the bioavailability of this Ag. Concentrations of Ag in the archived crop samples were generally <0.70 mg kg–1 in edible tissues, much less than those in diets that may cause an adverse effects in animals and humans (>100 mg kg–1). These data indicate that the transfer of Ag (derived from both traditional Ag industry and current nano Ag industry) to the terrestrial food chain is limited.

Arsenic (As) tends to mobilize in flooded paddy soil due to the reductive dissolution of the iron (oxyhydr)oxides to which As sorbs, resulting in elevated As accumulation in rice that poses a potential risk to the food safety and human health. Microbial sulfate reduction is an important biogeochemical process in paddy soils, but its impact on As mobilization remains poorly understood. In this study, we incubated eight As-contaminated paddy soils under flooded conditions to investigate the effect of sulfate addition on As mobility. Porewater Fe and As concentrations and As species were determined. Among the eight soils, an addition of 50 mg S kg−1 as sodium sulfate decreased porewater arsenite only in two soils, which also showed a high mobilization of Fe2+. Further experiments showed that the addition of sulfate to these two soils stimulated microbial sulfate reduction but decreased porewater concentrations of both arsenite and Fe2+. Additionally, the supply of sulfate increased the fractions of As associated with acid volatile sulfides in the solid phase and decreased As uptake by rice in pot experiments under similar conditions. The effect of sulfate addition on porewater As was diminished by the addition of molybdate, an inhibitor of sulfate reducing bacteria. These results suggest the formation of secondary FeS minerals which co-precipitate or sorb arsenite as a likely mechanism of As immobilization, which was also supported by thermodynamic modeling of the pore water. Thus, sulfate additions can immobilize As and reduce its availability to rice plants in paddy soils containing a high potential for microbial Fe reduction, providing an efficient way to mitigate the As transfer to the food chain.

Rice (Oryza sativa) is a major dietary source of the toxic metal cadmium (Cd). Reducing Cd transfer from soil to the rice grain is important for food safety. Rice cultivars vary widely in their Cd accumulation, but the genetic basis for this variation is not fully understood. Based on field and pot experiments comparing 26 rice cultivars, we identified a cultivar with high Cd accumulation in grain (BG367, coded as W4) and a cultivar with low grain Cd accumulation (Huajingxian 74, coded as W0). W4 showed a higher Cd translocation from roots to shoots than W0. Using chromosome single segment substitution lines derived from the two cultivars, we mapped a quantitative trait locus for Cd accumulation in grain to a 400 kb region in chromosome 7. Using yeast expression assays and transgenic complementation, we identified OsHMA3 as the causal gene at this locus. Compared with OsHMA3W0, OsHMA3W4 has a deletion of 14 amino acids predicted to be in the ATP binding domain. OsHMA3W4 showed a complete loss of transport activity for Cd in yeast assays. Taking our findings together, we have identified a new allele of OsHMA3 with a total loss-of-function, resulting in greatly elevated Cd translocation to rice shoots and grain.